Measuring pH of the Coxiella burnetii Parasitophorous Vacuole. Issue 1 (9th November 2017)

• Record Type: Journal Article
• Authors: Samanta, Dhritiman; Gilk, Stacey D.
• Editors: Coico, Richard; McBride, Alison; Quarles, John M.; Stevenson, Brian; Taylor, Ronald K.
• Is Part Of: Current protocols in microbiology. Volume 47:Issue 1(2017)
• Page Start: 6C.3.1
• Page End: 6C.3.11
• Publication Date: 2017-11-09
• Subjects: Coxiella burnetii -- parasitophorous vacuole -- pH -- quantitative microscopy; Microbiology -- Laboratory manuals; Microbiology -- Technique -- Periodicals; Microbiological Techniques -- methods; Containment of Biohazards -- methods; Laboratory Infection -- prevention & control; Microbiology; Microbiology -- Technique; Fulltext; Internet Resources; Laboratory Manuals; Reference Materials; Periodicals; Laboratory manuals; 579.028
• DOI: 10.1002/cpmc.38
• Languages: English
• ISSNs: 1934-8525

Abstract

Abstract: Coxiella burnetii is the causative agent of human Q fever, a zoonotic disease that can cause a debilitating, flu‐like illness in acute cases, or a life‐threatening endocarditis in chronic patients. An obligate intracellular bacterial pathogen, Coxiella survives and multiplies in a large lysosome‐like vacuole known as the Coxiella parasitophorous vacuole (CPV). A unique characteristic of the CPV is the acidic environment (pH ∼5.0), which is required to activate Coxiella metabolism and the Coxiella type 4 secretion system (T4SS), a major virulence factor required for intracellular survival. Further, inhibiting or depleting vacuolar ATPase, a host cell protein that regulates lysosomal pH, inhibits intracellular Coxiella growth. Together, these data suggest that CPV pH is an important limiting factor for Coxiella growth and virulence. This unit describes a method to determine CPV pH using live cell microscopy of a pH–sensitive fluorophore conjugated to dextran. This technique is useful to measure changes in CPV pH during infection or in response to drug treatment. © 2017 by John Wiley & Sons, Inc.